Inbred maize line PHBE2

ABSTRACT

An inbred maize line, designated PHBE2, the plants and seeds of inbred maize line PHBE2, methods for producing a maize plant produced by crossing the inbred line PHBE2 with itself or with another maize plant, and hybrid maize seeds and plants produced by crossing the inbred line PHBE2 with another maize line or plant.

FIELD OF THE INVENTION

This invention is in the field of maize breeding, specifically relating to an inbred maize line designated PHBE2.

BACKGROUND OF THE INVENTION

The goal of plant breeding is to combine in a single variety or hybrid various desirable traits. For field crops, these traits may include resistance to diseases and insects, tolerance to heat and drought, reducing the time to crop maturity, greater yield, and better agronomic quality. With mechanical harvesting of many crops, uniformity of plant characteristics such as germination and stand establishment, growth rate, maturity, and plant and ear height, is important.

Field crops are bred through techniques that take advantage of the plant's method of pollination. A plant is self-pollinated if pollen from one flower is transferred to the same or another flower of the same plant. A plant is cross-pollinated if the pollen comes from a flower on a different plant.

Plants that have been self-pollinated and selected for type for many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny. A cross between two different homozygous lines produces a uniform population of hybrid plants that may be heterozygous for many gene loci. A cross of two plants each heterozygous at a number of gene loci will produce a population of hybrid plants that differ genetically and will not be uniform.

Maize plants (zea mays L.), often referred to as corn in the United States, can be bred by both self-pollination and cross-pollination techniques. Maize has separate male and female flowers on the same plant, located on the tassel and the ear, respectively. Natural pollination occurs in maize when wind blows pollen from the tassels to the silks that protrude from the tops of the ears.

A reliable method of controlling male fertility in plants offers the opportunity for improved plant breeding. This is especially true for development of maize hybrids, that relies upon some sort of male sterility system. There are several options for controlling male fertility available to breeders, such as: manual or mechanical emasculation (or detasseling), cytoplasmic male sterility, genetic male sterility, gametocides and the like.

Hybrid maize seed is typically produced by a male sterility system incorporating manual or mechanical detasseling. Alternate strips of two inbred varieties of maize are planted in a field, and the pollen-bearing tassels are removed from one of the inbreds (female). Providing that there is sufficient isolation from sources of foreign maize pollen, the ears of the detasseled inbred is will be fertilized only from the other inbred (male), and the resulting seed is therefore hybrid and will form hybrid plants.

The laborious, and occasionally unreliable, detasseling process can be avoided by using cytoplasmic male-sterile (CMS) inbreds. Plants of a CMS inbred are male sterile as a result of factors resulting from the cytoplasmic, as opposed to the nuclear, genome. Thus, this characteristic is inherited exclusively through the female parent in maize plants, since only the female provides cytoplasm to the fertilized seed. CMS plants are fertilized with pollen from another inbred that is not male-sterile. Pollen from the second inbred may or may not contribute genes that make the hybrid plants male-fertile. Usually seed from detasseled fertile maize and CMS produced seed of the same hybrid are blended to insure that adequate pollen loads are available for fertilization when the hybrid plants are grown.

There are several methods of conferring genetic male sterility available, such as multiple mutant genes at separate locations within the genome that confer male sterility, as disclosed in U.S. Pat. Nos. 4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations as described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. These and all patents referred to are incorporated herein by reference. In addition to these methods, Albertsen et al., of Pioneer Hi-Bred, U.S. patent application Ser. No. 07/848,433, have developed a system of nuclear male sterility that includes: identifying a gene that is critical to male fertility; silencing this native gene that is critical to male fertility; removing the native promoter from the essential male fertility gene and replacing it with an inducible promoter; inserting this genetically engineered gene back into the plant; and thus creating a plant that is male sterile because the inducible promoter is not "on" resulting in the male fertility gene not being transcribed. Fertility is restored by inducing, or turning "on", the promoter, that in turn allows the gene that confers male fertility to be transcribed.

There are many other methods of conferring genetic male sterility in the art, each with it's own benefits and drawbacks. These methods use a variety of approaches such as delivering into the plant a gene encoding a cytotoxic substance associated with a male tissue specific promoter or an antisense system in which a gene critical to fertility is identified and an antisense to that gene is inserted in the plant (see: Fabinjanski, et al. EPO 89/3010153.8 publication no. 329,308 and PCT application PCT/CA90/00037 published as WO 90/08828).

Another system useful in controlling male sterility makes use of gametocides. Gametocides are not a genetic system, but rather a topical application of chemicals. These chemicals affect cells that are critical to male fertility. The application of these chemicals affects fertility in the plants only for the growing season in which the gametocide is applied (see Carlson, Glenn R., U.S. Pat. No.: 4,936,904). Application of the gametocide, timing of the application and genotype specificity often limit the usefulness of the approach.

The use of male sterile inbreds is but one factor in the development of maize hybrids. The development of maize hybrids requires, in general, the development of homozygous inbred lines, the crossing of these lines, and the evaluation of the crosses. Pedigree breeding and recurrent selection breeding methods are used to develop inbred lines from breeding populations. Breeding programs combine the genetic backgrounds from two or more inbred lines or various other broad-based sources into breeding pools from which new inbred lines are developed by selfing and selection of desired phenotypes. The new inbreds are crossed with other inbred lines and the hybrids from these crosses are evaluated to determine which of those have commercial potential.

Pedigree breeding starts with the crossing of two genotypes, each of that may have one or more desirable characteristics that is lacking in the other or that complement the other. If the two original parents do not provide all the desired characteristics, other sources can be included in the breeding population. In the pedigree method, superior plants are selfed and selected in successive generations. In the succeeding generations the heterozygous condition gives way to homogeneous lines as a result of self-pollination and selection. Typically in the pedigree method of breeding five or more generations of selfing and selection is practiced: F₁ →F₂ ;F₃ .increment.F₄ ; F₄ →F₅, etc.

Recurrent selection breeding, backcrossing for example, can be used to improve an inbred line. Backcrossing can be used to transfer a specific desirable trait from one inbred or source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (recurrent parent) to a donor inbred (non-recurrent parent), that carries the appropriate gene(s) for the trait in question. The progeny of this cross is then mated back to the superior recurrent parent followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny will be heterozygous for loci controlling the characteristic being transferred, but will be like the superior parent for most or almost all other genes. The last backcross generation would be selfed to give pure breeding progeny for the gene(s) being transferred.

A single cross hybrid maize variety is the cross of two inbred lines, each of which has a genotype that complements the genotype of the other. The hybrid progeny of the first generation is designated F₁. In the development of hybrids only the F₁ hybrid plants are sought. Preferred F₁ hybrids are more vigorous than their inbred parents. This hybrid vigor, or heterosis, can be manifested in many polygenic traits, including increased vegetative growth and increased yield.

The development of a hybrid maize variety involves three steps:

(1) the selection of plants from various germplasm pools for initial breeding crosses;

(2) the selfing of the selected plants from the breeding crosses for several generations to produce a series of inbred lines, that, although different from each other, breed true and are highly uniform; and

(3) crossing the selected inbred lines with different inbred lines to produce the hybrid progeny (F₁).

During the inbreeding process in maize, the vigor of the lines decreases. Vigor is restored when two different inbred lines are crossed to produce the hybrid progeny (F₁). An important consequence of the homozygosity and homogeneity of the inbred lines is that the hybrid between any two inbreds will always be the same. Once the inbreds that give a superior hybrid have been identified, the hybrid seed can be reproduced indefinitely as long as the homogeneity of the inbred parents is maintained.

A single cross hybrid is produced when two inbred lines are crossed to produce the F₁ progeny. A double cross hybrid is produced from four inbred lines crossed in pairs (A×B and C×D) and then the two F₁ hybrids are crossed again (A×B)×(C×D). Much of the hybrid vigor exhibited by F₁ hybrids is lost in the next generation (F₂). Consequently, seed from hybrid varieties is not used for planting stock.

Maize is an important and valuable field crop. Thus, a continuing goal of plant breeders is to develop high-yielding maize hybrids that are agronomically sound based on stable inbred lines. The reasons for this goal are obvious: to maximize the amount of grain produced with the inputs used and minimize susceptibility of the crop to environmental stresses. To accomplish this goal, the maize breeder must select and develop superior inbred parental lines for producing hybrids. This requires identification and selection of genetically unique individuals that occur in a segregating population. The segregating population is the result of a combination of crossover events plus the independent assortment of specific combinations of alleles at many gene loci that results in specific genotypes. Based on the number of segregating genes, the frequency of occurrence of an individual with a specific genotypes. Based on the number of segregating genes, the frequency of occurrence of an individual with a specific genotype is less than 1 in 10,000. Thus, even if the entire genotype of the parents has been characterized and the desired genotype is known, only a few if any individuals having the desired genotype may be found in a large F₂ or S₀ population. Typically, however, the genotype of neither the parents nor the desired genotype is known in any detail.

In addition to the preceding problem, it is not known how the genotype will react with the environment. This genotype by environment interaction is an important, yet unpredictable, factor in plant breeding. A breeder of ordinary skill in the art cannot predict the genotype, how that genotype will interact with various environments or the resulting phenotypes of the developing lines, except perhaps in a very broad and general fashion. A breeder of ordinary skill in the art would also be unable to recreate the same line twice from the very same original parents as the breeder is unable to direct how the genomes combine or how they will interact with the environmental conditions. This unpredictability results in the expenditure of large-amounts of research resources in the development of a superior new maize inbred line.

Pioneer research station staff propose about 400 to 500 new inbreds each year from over 2,000,000 pollinations. Of those proposed new inbreds, less than 50 and more commonly less than 30 are actually selected for commercial use.

Summary of the Invention

According to the invention, there is provided a novel inbred maize line, designated PHBE2. This invention thus relates to the seeds of inbred maize line PHBE2, to the plants of inbred maize line PHBE2, and to methods for producing a maize plant produced by crossing the inbred line PHBE2 with itself or another maize line. This invention further relates to hybrid maize seeds and plants produced by crossing the inbred line PHBE2 with another maize line.

Definitions

In the description and examples that follow, a number of terms are used herein. In order to provide a clear and consistent understanding of the specification and claims, including the scope to be given such terms, the following definitions are provided. NOTE: ABS is in absolute terms and %MN is percent of the mean for the experiments in which the inbred or hybrid was grown. These designators will follow the descriptors to denote how the values are to be interpreted. Below are the descriptors used in the data tables included herein.

ANT ROT=ANTHRACNOSE STALK ROT (Colletotrichum graminicola). A 1 to 9 visual rating indicating the resistance to Anthracnose Stalk Rot. A higher score indicates a higher resistance.

BAR PLT=BARREN PLANTS. The percent of plants per plot that were not barren (lack ears).

BRT STK=BRITTLE STALKS. This is a measure of the stalk breakage near the time of pollination, and is an indication of whether a hybrid or inbred would snap or break near the time of flowering under severe winds. Data are presented as percentage of plants that did not snap.

BU ACR=YIELD (BUSHELS/ACRE). Yield of the grain at harvest in bushels per acre adjusted to 15.5% moisture.

CLN=CORN LETHAL NECROSIS (synergistic interaction of maize chlorotic mottle virus (MCMV) in combination with either maize dwarf mosaic virus (MDMV-A or MDMV-B) or wheat streak mosaic virus (WSMV)). A 1 to 9 visual rating indicating the resistance to Corn Lethal Necrosis. A higher score indicates a higher resistance.

COM RST=COMMON RUST (Puccinia sorghi). A 1 to 9 visual rating indicating the resistance to Common Rust. A higher score indicates a higher resistance.

D/D=DRYDOWN. This represents the relative rate at which a hybrid will reach acceptable harvest moisture compared to other hybrids on a 1-9 rating scale. A high score indicates a hybrid that dries relatively fast while a low score indicates a hybrid that dries slowly.

DIP ERS=DIPLODIA EAR MOLD SCORES (Diplodia maydis and Diplodia macrospora). A 1 to 9 visual rating indicating the resistance to Diplodia Ear Mold. A higher score indicates a higher resistance.

DRP EAR=DROPPED EARS. A measure of the number of dropped ears per plot and represents the percentage of plants that did not drop ears prior to harvest.

D/T=DROUGHT TOLERANCE. This represents a 1-9 rating for drought tolerance, and is based on data obtained under stress conditions. A high score indicates good drought tolerance and a low score indicates poor drought tolerance.

EAR HT=EAR HEIGHT. The ear height is a measure from the ground to the highest placed developed ear node attachment and is measured in inches.

EAR MLD=General Ear Mold. Visual rating (1-9 score) where a "1" is very susceptible and a "9" is very resistant. This is based on overall rating for ear mold of mature ears without determining the specific mold organism, and may not be predictive for a specific ear mold.

EAR SZ=EAR SIZE. A 1 to 9 visual rating of ear size. The higher the rating the larger the ear size.

ECB 1LF=EUROPEAN CORN BORER FIRST GENERATION LEAF FEEDING (Ostrinia nubilalis). A 1 to 9 visual rating indicating the resistance to preflowering leaf feeding by first generation European Corn Borer. A higher score indicates a higher resistance.

ECB 2IT=EUROPEAN CORN BORER SECOND GENERATION INCHES OF TUNNELING (Ostrinia nubilalis). Average inches of tunneling per plant in the stalk.

ECB 2SC=EUROPEAN CORN BORER SECOND GENERATION (Ostrinia nubilalis). A 1 to 9 visual rating indicating post flowering degree of stalk breakage and other evidence of feeding by European Corn Borer, Second Generation. A higher score indicates a higher resistance.

ECB DPE=EUROPEAN CORN BORER DROPPED EARS (Ostrinia nubilalis). Dropped ears due to European Corn Borer. Percentage of plants that did not drop ears under second generation corn borer infestation.

EST CNT=EARLY STAND COUNT. This is a measure of the stand establishment in the spring and represents the number of plants that emerge on per plot basis for the inbred or hybrid.

EYE SPT=Eye Spot (Kabatiella zeae or Aureobasidiurn zeae). A 1 to 9 visual rating indicating the resistance to Eye Spot. A higher score indicates a higher resistance.

FUS ERS=FUSARIUM EAR ROT SCORE (Fusarium moniliforme or Fusarium subglutinans). A 1 to 9 visual rating indicating the resistance to Fusarium ear rot. A higher score indicates a higher resistance.

GDU=Growing Degree Units. Using the Barger Heat Unit Theory, that assumes that maize growth occurs in the temperature range 50° F.-86° F. and that temperatures outside this range slow down growth; the maximum daily heat unit accumulation is 36 and the minimum daily heat unit accumulation is 0. The seasonal accumulation of GDU is a major factor in determining maturity zones.

GDU SHD=GDU TO SHED. The number of growing degree units (GDUs) or heat units required for an inbred line or hybrid to have approximately 50 percent of the plants shedding pollen and is measured from the time of planting. Growing degree units are calculated by the Barger Method, where the heat units for a 24-hour period are: ##EQU1##

The highest maximum temperature used is 86° F. and the lowest minimum temperature used is 50° F. For each inbred or hybrid it takes a certain number of GDUs to reach various stages of plant development.

GDU SLK=GDU TO SILK. The number of growing degree units required for an inbred line or hybrid to have approximately 50 percent of the plants with silk emergence from time of planting. Growing degree units are calculated by the Barger Method as given in GDU SHD definition.

GIB ERS=GIBBERELLA EAR ROT (PINK MOLD) (Gibberella zeae). A 1 to 9 visual rating indicating the resistance to Gibberella Ear Rot. A higher score indicates a higher resistance.

GLF SPT=Gray Leaf Spot (Cercospora zeae-maydis). A 1 to 9 visual rating indicating the resistance to Gray Leaf Spot. A higher score indicates a higher resistance.

GOS WLT=Goss' Wilt (Corynebacterium nebraskense). A 1 to 9 visual rating indicating the resistance to Goss' Wilt. A higher score indicates a higher resistance.

GRN APP=GRAIN APPEARANCE. This is a 1 to 9 rating for the general appearance of the shelled grain as it is harvested based on such factors as the color of harvested grain, any mold on the grain, and any cracked grain. High scores indicate good grain quality.

H/POP=YIELD AT HIGH DENSITY. Yield ability at relatively high plant densities on 1-9 relative rating system with a higher number indicating the hybrid responds well to high plant densities for yield relative to other hybrids. A 1, 5, and 9 would represent very poor, average, and very good yield response, respectively, to increased plant density.

HC BLT=HELMINTHOSPORIUM CARBONUM LEAF BLIGHT (Helminthosporium carbonum). A 1 to 9 visual rating indicating the resistance to Helminthosporium infection. A higher score indicates a higher resistance.

HD SMT=HEAD SMUT (Sphacelotheca reiliana). This score indicates the percentage of plants not infected.

INC D/A=GROSS INCOME (DOLLARS PER ACRE). Relative income per acre assuming drying costs of two cents per point above 15.5 percent harvest moisture and current market price per bushel.

INCOME/ACRE. Income advantage of hybrid to be patented over other hybrid on per acre basis.

INC ADV=GROSS INCOME ADVANTAGE. GROSS INCOME advantage of variety #1 over variety #2.

L/POP=YIELD AT LOW DENSITY. Yield ability at relatively low plant densities on a 1-9 relative system with a higher number indicating the hybrid responds well to low plant densities for yield relative to other hybrids. A 1, 5, and 9 would represent very poor, average, and very good yield response, respectively, to low plant density.

MDM CPX=MAIZE DWARF MOSAIC COMPLEX (MDMV=Maize Dwarf Mosaic Virus and MCDV=Maize Chlorotic Dwarf Virus). A 1 to 9 visual rating indicating the resistance to Maize Dwarf Mosaic Complex. A higher score indicates a higher resistance.

MST=HARVEST MOISTURE. The moisture is the actual percentage moisture of the grain at harvest.

MST ADV=MOISTURE ADVANTAGE. The moisture advantage of variety #1 over variety #2 as calculated by: MOISTURE of variety #2 MOISTURE of variety #1=MOISTURE ADVANTAGE of variety #1.

NLF BLT=Northern Leaf Blight (Helminthosporium turcicum or Exserohilum turcicum). A 1 to 9 visual rating indicating the resistance to Northern Leaf Blight. A higher score indicates a higher resistance.

PLT HT=PLANT HEIGHT. This is a measure of the height of the plant from the ground to the tip of the tassel in inches.

POL SC=POLLEN SCORE. A 1 to 9 visual rating indicating the amount of pollen shed. The higher the score the more pollen shed.

POL WT=POLLEN WEIGHT. This is calculated by dry weight of tassels collected as shedding commences minus dry weight from similar tassels harvested after shedding is complete.

It should be understood that the inbred can, through routine manipulation of cytoplasmic or other factors, be produced in a male-sterile form.

POP K/A=PLANT POPULATIONS. Measured as 1000s per acre.

POP ADV=PLANT POPULATION ADVANTAGE. The plant population advantage of variety #1 over variety #2 as calculated by PLANT POPULATION of variety #2 - PLANT POPULATION of variety #1=PLANT POPULATION ADVANTAGE of variety #1.

PRM=PREDICTED Relative Maturity. This trait, predicted relative maturity, is based on the harvest moisture of the grain. The relative maturity rating is based on a known set of checks and utilizes standard linear regression analyses and is referred to as the Comparative Relative Maturity Rating System that is similar to the Minnesota Relative Maturity Rating System.

PRM SHD=A relative measure of the growing degree units (GDU) required to reach 50% pollen shed. Relative values are predicted values from the linear regression of observed GDU's on relative maturity of commercial checks.

RT LDG=ROOT LODGING. Root lodging is the percentage of plants that do not root lodge; plants that lean from the vertical axis at an approximately 30° angle or greater would be counted as root lodged.

RTL ADV=ROOT LODGING ADVANTAGE. The root lodging advantage of variety #1 over variety #2.

SCT GRN=SCATTER GRAIN. A 1 to 9 visual rating indicating the amount of scatter grain (lack of pollination or kernel abortion) on the ear. The higher the score the less scatter grain.

SDG VGR=SEEDLING VIGOR. This is the visual rating (1 to 9) of the amount of vegetative growth after emergence at the seedling stage (approximately five leaves). A higher score indicates better vigor.

SEL IND=SELECTION INDEX. The selection index gives a single measure of the hybrid's worth based on information for up to five traits. A maize breeder may utilize his or her own set of traits for the selection index. One of the traits that is almost always included is yield. The selection index data presented in the tables represent the mean value averaged across testing stations.

SLF BLT=SOUTHERN LEAF BLIGHT (Helminthosporium maydis or Bipolaris maydis). A 1 to 9 visual rating indicating the resistance to Southern Leaf Blight. A higher score indicates a higher resistance.

SOU RST=SOUTHERN RUST (Puccinia polysora). A 1 to 9 visual rating indicating the resistance to Southern Rust. A higher score indicates a higher resistance.

STA GRN=STAY GREEN. Stay green is the measure of plant health near the time of black layer formation (physiological maturity). A high score indicates better late-season plant health.

STD ADV=STALK STANDING ADVANTAGE. The advantage of variety #1 over variety #2 for the trait STK CNT.

STK CNT=NUMBER OF PLANTS. This is the final stand or number of plants per plot.

STK LDG=STALK LODGING. This is the percentage of plants that did not stalk lodge (stalk breakage) as measured by either natural lodging or pushing the stalks and determining the percent/age of plants that break below the ear.

STW WLT=Stewart's Wilt (Erwinia stewartii). A 1 to 9 visual rating indicating the resistance to Stewart's Wilt. A higher score indicates a higher resistance.

TAS BLS=TASSEL BLAST. A 1 to 9 visual rating was used to measure the degree of blasting (necrosis due to heat stress) of the tassel at the time of flowering. A 1 would indicate a very high level of blasting at time of flowering, while a 9 would have no tassel blasting.

TAS SZ=TASSEL SIZE. A 1 to 9 visual rating was used to indicate the relative size of the tassel. The higher the rating the larger the tassel.

TAS WT=TASSEL WEIGHT. This is the average weight of a tassel (grams) just prior to pollen shed.

TEX EAR=EAR TEXTURE. A 1 to 9 visual rating was used to indicate the relative hardness (smoothness of crown) of mature grain. A 1 would be very soft (extreme dent) while a 9 would be very hard (flinty or very smooth crown).

TILLER=TILLERS. A count of the number of tillers per plot that could possibly shed pollen was taken. Data is given as a percentage of tillers: number of tillers per plot divided by number of plants per plot.

TST WT=TEST WEIGHT (UNADJUSTED). The measure of the weight of the grain in pounds for a given volume (bushel).

TST WTA=TEST WEIGHT ADJUSTED. The measure of the weight of the grain in pounds for a given volume (bushel) adjusted for 15.5 percent moisture.

TSW ADV=TEST WEIGHT ADVANTAGE. The test weight advantage of variety #1 over variety #2.

WIN M%=PERCENT MOISTURE WINS.

WIN Y%=PERCENT YIELD WINS.

YLD=YIELD. It is the same as BU ACR ABS.

YLD ADV=YIELD ADVANTAGE. The yield advantage of variety #1 over variety #2 as calculated by: YIELD of variety #1 - YIELD variety #2=yield advantage of variety #1.

YLD SC=YIELD SCORE. A 1 to 9 visual rating was used to give a relative rating for yield based on plot ear piles. The higher the rating the greater visual yield appearance.

DETAILED DESCRIPTION OF THE INVENTION

Inbred maize lines are typically developed for use in the production of hybrid maize lines. Inbred maize lines need to be highly homogeneous, homozygous and reproducible to be useful as parents of commercial hybrids. There are many analytical methods available to determine the homozygotic and phenotypic stability of these inbred lines.

The oldest and most traditional method of analysis is the observation of phenotypic traits. The data is usually collected in field experiments over the life of the maize plants to be examined. Phenotypic characteristics most often observed are for traits associated with plant morphology, ear and kernel morphology, insect and disease resistance, maturity, and yield.

In addition to phenotypic observations, the genotype of a plant can also be examined. There are many laboratory-based techniques available for the analysis, comparison and characterization of plant genotype; among these are Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), and Simple Sequence Repeats (SSRs) which are also referred to as Microsatellites.

The most widely used of these laboratory techniques are Isozyme Electrophoresis and RFLPs as discussed in Lee, M., "Inbred Lines of Maize and Their Molecular Markers," The Maize Handbook, (Springer-Verlag, New York, Inc. 1994, at 423-432) incorporated herein by reference. Isozyme Electrophoresis is a useful tool in determining genetic composition, although it has relatively low number of available markers and the low number of allelic variants among maize inbreds. RFLPs have the advantage of revealing an exceptionally high degree of allelic variation in maize and the number of available markers is almost limitless.

Maize RFLP linkage maps have been rapidly constructed and widely implemented in genetic studies. One such study is described in Boppenmaier, et al., "Comparisons among strains of inbreds for RFLPs", Maize Genetics Cooperative Newsletter, 65:1991, pg. 90, is incorporated herein by reference. This study used 101 RFLP markers to analyze the patterns of 2 to 3 different deposits each of five different inbred lines. The inbred lines had been selfed from 9 to 12 times before being adopted into 2 to 3 different breeding programs. It was results from these 2 to 3 different breeding programs that supplied the different deposits for analysis. These five lines were maintained in the separate breeding programs by selfing or sibbing and rogueing off-type plants for an additional one to eight generations. After the RFLP analysis was completed, it was determined the five lines showed 0-2% residual heterozygosity. Although this was a relatively small study, it can be seen using RFLPs that the lines had been highly homozygous prior to the separate strain maintenance.

Inbred maize line PHBE2 produces high yields in hybrid combinations and is a very good male in production fields. Inbred maize line PHBE2 is a yellow, dent maize inbred that provides an acceptable male parental line in crosses for producing first generation F1 maize hybrids. Inbred PHBE2 is adapted to the Southwest region of the United States. Inbred PHBE2 tends to produce hybrids with below average resistance to Brittle Stalk.

The inbred has shown uniformity and stability within the limits of environmental influence for all the traits as described in the Variety Description Information (Table 1) that follows. Most of the data in the Variety Description information was collected at Johnston, Iowa. The inbred has been self-pollinated and ear-rowed a sufficient number of generations with careful attention paid to uniformity of plant type to ensure the homozygosity and phenotypic stability necessary to use in commercial production. The line has been increased both by hand and in isolated fields with continued observation for uniformity. No variant traits have been observed or are expected in PHBE2.

Inbred maize line PHBE2, being substantially homozygous, can be reproduced by planting seeds of the line, growing the resulting maize plants under self-pollinating or sib-pollinating conditions with adequate isolation, and harvesting the resulting seed, using techniques familiar to the agricultural arts.

                  TABLE 1                                                          ______________________________________                                         VARIETY DESCRIPTION INFORMATION                                                INBRED = PHBE2                                                                 ______________________________________                                         Type: Dent    Region Best Adapted: Southwest                                   A.    Maturity: Average across maturity zones. Zone: 0                               GDU Shed: 1490                                                                 GDU Silk: 1520                                                                 No. Reps: 47                                                             B.    Plant Characteristics:                                                         Plant height (to tassel tip): 233 cm                                           Length of top ear internode: 15 cm                                             Number of ears per stalk: Slight two-ear tendency                              Ear height (to base of top ear): 84 cm                                         Number of tillers: None                                                        Cytoplasm type: Normal                                                   C.    Leaf:                                                                          Color: (B14) Dark Green*                                                       Angle from Stalk: <30 degrees                                                  Marginal Waves: (WF9) Few                                                      Sheath Pubescence: (W22) Light                                                 Longitudinal Creases: (OH56A) Few                                              Length (Ear node leaf): 73 cm                                                  Width (widest point, ear node leaf): 9 cm                                D.    Tassel:                                                                        Number lateral branches: 12                                                    Branch Angle from central spike: 30-40 degrees                                 Pollen Shed: (KY21) Heavy based on Pollen Yield Test                           (134% of experiment means)                                                     Peduncle Length (top leaf to basal branches): 21 cm                            Anther Color: Yellow*                                                          Glume Color: Green*                                                      E.    Ear (Husked Ear Data Except When Stated Otherwise):                            Length: 15 cm                                                                  Weight: 108 gm                                                                 Mid-point Diameter: 44 mm                                                      Silk Color: Pink*                                                              Husk Extension (Harvest stage): Short (Ears) Exposed                           Husk Leaf: Short (L8 cm)                                                       Taper of Ear: Average                                                          Position of Shank (dry husks): pendent                                         Kernel Rows: Straight, distinct Number = 16                                    Husk Color (fresh): Light Green*                                               Husk Color (dry): Buff*                                                        Shank Length: 9 cm                                                             Shank (No. of internodes): 7                                             F.    Kernel (Dried):                                                                Size (from ear mid-point)                                                      Length: 11 mm                                                                  Width: 9 mm                                                                    Thick: 4 mm                                                                    Pericarp Color: Colorless*                                                     Aleurone Color: Homozygous Yellow*                                             Endosperm Color: Yellow*                                                       Endosperm Type: Normal                                                         Gm Wt/100 Seeds (unsized): 23 gm                                         G.    Cob:                                                                           Diameter at mid-point: 27 mm                                                   Strength: Strong                                                               Color: Red*                                                              H.    Diseases:                                                                      Corn Lethal Necrosis (MCMV = Maize Chlorotic                                   Mottle Virus and MDMV = Maize Dwarf Mosaic                                     Virus): Intermediate                                                           Maize Dwarf Mosaic Complex (MDMV & MCDV =                                      Maize Dwarf Virus): Intermediate                                               Anthracnose Stalk Rot (C. graminicola): Susceptible                            N. Leaf Blight (E. turcicum): Intermediate                                     Common Rust (P. sorghi): Intermediate                                          Southern Rust (P. polysora): Susceptible                                       Gray Leaf Spot (C. zeae): Susceptible                                          Stewart's Wilt (E. stewartii): Resistant                                       Head Smut (S. reiliana): Resistant                                             Fusarium Ear Mold (F. moniliforme): Intermediate                         I.    Insects:                                                                       European Corn Borer-1 Leaf Damage (Pre-flowering):                             Resistant                                                                      European Corn Borer-2 (Post-flowering): Susceptible                            The above descriptions are based on a scale of 1-9,                            1 being highly susceptible, 9 beeing highly resistant.                         S (Susceptible): Would generally represent a score of                          1-3.                                                                           I (intermediate): Would generally represent a score of                         4-5.                                                                           R (Resistant): Would generally represent a score of 6-7.                       H (Highly Resistant): Would generally rpresent a score                         of 8-9.                                                                        Highly resistant does not imply the inbred is                                  immune.                                                                  J.    Variety Most Closely Resembling:                                               Character Inbred                                                               Maturity PHN46                                                                 Usage PHN46                                                              ______________________________________                                          *In interpreting the foregoing color designations, reference may be had t      the Munsell Glossy Book of Color, a standard color reference.                  Data for Items B, C, D, E, F, and G is based primarily on a maximum of 4       reps from Johnston, Iowa grown in 1993 and 1994, plus description              information from the maintaining station.                                      PHN46 (PVP Certificate No. 9000249) is a Pioneer HiBred International,         Inc. proprietary inbred.                                                 

ELECTROPHORESIS RESULTS Isozyme Genotypes for PHBE2

Isozyme data were generated for inbred maize line PHBE2 according to the procedures described in Stuber, C. W., Wendel, J. F., Goodman, M. M., and Smith, J. S. C., "Techniques and Scoring Procedures for Starch Gel Electrophoresis of Enzymes from Maize (Zea mays L.)", Technical Bulletin No. 286, North Carolina Agricultural Research Service, North Carolina State University, Raleigh, N.C. (1988).

The data in Table 2 compares PHBE2 with its parents, PHR03 and PHN46.

                  TABLE 2                                                          ______________________________________                                         ELECTROPHORESIS RESULTS FOR PBHE2                                              AND ITS PARENTS PHR03 AND PHN46                                                       PARENTS                                                                 LOCI     PHBE2         PHR03   PHN46                                           ______________________________________                                         ACP1     4             4       3                                               ADH1     4             4       4                                               CAT3     9             9       9                                               DIA1     8             8       8                                               GOT1     4             4       4                                               GOT2     4             4       4                                               GOT3     4             4       4                                               IDH1     4             4       4                                               IDH2     6             6       6                                               MDH1     6             6       6                                               MDH2     3.5           3.5     3.5                                             MDH3     16            16      18                                              MDH4     12            12      12                                              MDH5     12            12      12                                              MMM      4             4       4                                               PGM1     9             9       9                                               PGM2     4             3       4                                               PGD1     3.8           3.8     2                                               PGD2     5             5       5                                               PHI1     4             4       4                                               ______________________________________                                    

Industrial Applicability

This invention also is directed to methods for producing a maize plant by crossing a first parent maize plant with a second parent maize plant wherein the first or second parent maize plant is an inbred maize plant from the line PHBE2. Further, both first and second parent maize plants can come from the inbred maize line PHBE2. Thus, any such methods using the inbred maize line PHBE2 are part of this invention: selfing, backcrosses, hybrid production, crosses to populations, and the like. All plants produced using inbred maize line PHBE2 as a parent are within the scope of this invention. Advantageously, the inbred maize line is used in crosses with other, different, maize inbreds to produce first generation (F₁) maize hybrid seeds and plants with superior characteristics.

As used herein, the term plant includes plant cells, plant protoplasts, plant cell tissue cultures from which maize plants can be regenerated, plant calli, plant clumps, and plant cells that are intact in plants or parts of plants, such as embryos, pollen, flowers, kernels, ears, cobs, leaves, husks, stalks, roots, root tips, anthers, silk and the like.

Duncan, Williams, Zehr, and Widholm, Planta (1985) 165:322-332 reflects that 97% of the plants cultured that produced callus were capable of plant regeneration. Subsequent experiments with both inbreds and hybrids produced 91% regenerable callus that produced plants. In a further study in 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988), 7:262-265 reports several media additions that enhance regenerability of callus of two inbred lines. Other published reports also indicated that "nontraditional" tissues are capable of producing somatic embryogenesis and plant regeneration. K. P. Rao, et al., Maize Genetics Cooperation Newsletter, 60:64-65 (1986), refers to somatic embryogenesis from glume callus cultures and B. V. Conger, et al., Plant Cell Reports, 6:345-347 (1987) indicates somatic embryogenesis from the tissue cultures of maize leaf segments. Thus, it is clear from the literature that the state of the art is such that these methods of obtaining plants are, and were, "conventional" in the sense that they are routinely used and have a very high rate of success.

Tissue culture of maize is described in European Patent Application, publication 160,390, incorporated herein by reference. Maize tissue culture procedures are also described in Green and Rhodes, "Plant Regeneration in Tissue Culture of Maize," Maize for Biological Research (Plant Molecular Biology Association, Charlottesville, Va. 1982, at 367-372) and in Duncan, et al., "The Production of Callus Capable of Plant Regeneration from Immature Embryos of Numerous Zea Mays Genotypes," 165 Planta 322-332 (1985). Thus, another aspect of this invention is to provide cells that upon growth and differentiation produce maize plants having the physiological and morphological characteristics of inbred line PHBE2.

Maize is used as human food, livestock feed, and as raw material in industry. The food uses of maize, in addition to human consumption of maize kernels, include both products of dry- and wet-milling industries. The principal products of maize dry milling are grits, meal and flour. The maize wet-milling industry can provide maize starch, maize syrups, and dextrose for food use. Maize oil is recovered from maize germ, that is a by-product of both dry- and wet-milling industries.

Maize, including both grain and non-grain portions of the plant, is also used extensively as livestock feed, primarily for beef cattle, dairy cattle, hogs, and poultry.

Industrial uses of maize include production of ethanol, maize starch in the wet-milling industry and maize flour in the dry-milling industry. The industrial applications of maize starch and flour are based on functional properties, such as viscosity, film formation, adhesive properties, and ability to suspend particles. The maize starch and flour have application in the paper and textile industries. Other industrial uses include applications in adhesives, building materials, foundry binders, laundry starches, explosives, oil-well muds, and other mining applications.

Plant parts other than the grain of maize are also used in industry: for example, stalks and husks are made into paper and wallboard and cobs are used for fuel and to make charcoal.

The seed of inbred maize line PHBE2, the plant produced from the inbred seed, the hybrid maize plant produced from the crossing of the inbred, hybrid seed, and various parts of the hybrid maize plant can be utilized for human food, livestock feed, and as a raw material in industry.

Performance Examples of PHBE2

In the examples that follow, the traits and characteristics of inbred maize line PHBE2 are given as a line in comparison with other inbreds and in hybrid combination. The data collected on inbred maize line PHBE2 is presented for the key characteristics and traits.

Table 3A compares PHBE2 to its PHR03 parent. The inbreds have similar yields and grain harvest moisture but PHBE2 is a shorter inbred with lower ear placement.

Table 3B compares PHBE2 to its PHN46 parent. PHBE2 and PHN46 have similar yield and grain harvest moisture but PHBE2 is taller with higher ear placement compared to PHN46.

Table 4A compares PHBE2 to PHR03 when both were crossed to the same inbreds and grown in the same experiments. The hybrids have similar yield and test weight but the PHBE2 hybrids have slightly lower grain harvest moisture compared to the PHR03 hybrids. The PHBE2 hybrids have better staygreen and seedling vigor than the PHR03 hybrids.

Table 4B compares PHBE2 to PHM46 when both were crossed to the same inbreds and grown in the same experiments. The hybrids have similar yield and agronomic characteristics but the PHBE2 hybrids are taller with higher ear placement compared to the PHM46 hybrids.

                                      TABLE 3A                                     __________________________________________________________________________     PAIRED INBRED COMPARISON DATA                                                  VARIETY #1 = PHBE2                                                             VARIETY #2 = PHR03                                                             __________________________________________________________________________                     BU  BU      STK RT  STA TST                                            VAR PRM ACR ACR MST LDG LDG GRN WTA                                            #   SHD ABS \% MN                                                                              \% MN                                                                              \% MN                                                                              \% MN                                                                              \% MN                                                                              \% MN                                  __________________________________________________________________________     TOTAL SUM                                                                              1   111 60.8                                                                               112 100 90  102 78  103                                            2   113 65.5                                                                               120 106 92  86  115 101                                            LOCS                                                                               4   4   4   4   2   3   26  2                                              REPS                                                                               4   8   8   8   4   3                                                      DIFF                                                                               2   4.7 8   6   2   16  37  2                                              PROB                                                                               .027+                                                                              .432                                                                               .456                                                                               .363                                                                               .849                                                                               .658                                                                               .000#                                                                              .530                                   __________________________________________________________________________                 SDG  EST STK  PLT EAR  BRT GDU                                             VAR VGR  CNT CNT  HT  HT   STK SHD                                             #   % MN % MN                                                                               % MN % MN                                                                               % MN % MN                                                                               % MN                                    __________________________________________________________________________     TOTAL SUM                                                                              1   95   95  99   101 103  102 101                                             2   100  106 106  108 111  107 104                                             LOCS                                                                               57   90  66   29  29   19  71                                              REPS                                                                               65   152 89   33  33   23  91                                              DIFF                                                                               4    11  7    7   8    5   3                                               PROB                                                                               .333 .000#                                                                              .005#                                                                               .000#                                                                              .000#                                                                               .023+                                                                              .000#                                   __________________________________________________________________________      *10% SIG                                                                       + = 5% SIG                                                                     # = 1% SIG                                                               

                                      TABLE 3B                                     __________________________________________________________________________     PAIRED INBRED COMPARISON DATA                                                  VARIETY #1 = PHBE2                                                             VARIETY #2 = PHN46                                                             __________________________________________________________________________                     BU  BU      STK RT  STA TST                                            VAR PRM ACR ACR MST LDG LDG GRN WTA                                            #   SHD ABS \% MN                                                                              \% MN                                                                              \% MN                                                                              \% MN                                                                              \% MN                                                                              \% MN                                  __________________________________________________________________________     TOTAL SUM                                                                              1   111 55.6                                                                               109 87  90  100 77  103                                            2   110 52.9                                                                               103 91  105 107 73  96                                             LOCS                                                                               4   2   2   2   2   4   23  2                                              REPS                                                                               4   4   4   4   4   6   27  DIFF 1 2.7 6 5 15 8 5 7                        PROB                                                                               .222                                                                               .646                                                                               .619                                                                               .686                                                                               .470                                                                               .000#                                                                              .542                                                                               .280                                   __________________________________________________________________________                 SDG  EST STK  PLT EAR  BRT GDU                                             VAR VGR  CNT CNT  HT  HT   STK SHD                                             #   % MN % MN                                                                               % MN % MN                                                                               % MN % MN                                                                               % MN                                    __________________________________________________________________________     TOTAL SUM                                                                              1   95   93  97   101 103  100 101                                             2   97   101 104  95  95   102 101                                             LOCS                                                                               57   96  73   30  30   20  83                                              REPS                                                                               63   164 103  34  34   24  111                                             DIFF                                                                               3    7   6    6   8    2   0                                               PROB                                                                               .557 .003#                                                                              .003#                                                                               .001#                                                                              .002#                                                                               .385                                                                               .419                                    __________________________________________________________________________      *10% SIG                                                                       + = 5% SIG                                                                     # = 1% SIG                                                               

                                      TABLE 4A                                     __________________________________________________________________________     Average Inbred By Tester Performance Comparing PHBE2 to PHR03 Crossed To       The Same                                                                       Inbred Testers And Grown in The Same Experiments. All Values Are               Expressed As Percent Of                                                        The Experiment Mean Except Predicted RM, Selection Index, and Yield            (BU/ACR)                                                                                        SEL BU          GDU PRM STK RT  STA                                  INBRED                                                                               PRM IND ACR YLD MST SHD SHD LDG LDG GRN                           __________________________________________________________________________     TOTAL  REPLIC.                                                                              132 132 141 141 142 37  27  107 47  61                            MEAN WTS                                                                              PHBE2 112 100 143 99  97  102 114 98  100 110                           MEAN WTS                                                                              PHR03 113 102 145 101 100 103 115 99  99  105                                  DIFF  1   2   2   2   4   0   0   1   1   6                             __________________________________________________________________________                  TST  GRN SDG EST STK  PLT EAR  DRP BRT                                   INBRED                                                                               WTA  APP VGR CNT CNT  HT  HT   EAR STK                            __________________________________________________________________________     TOTAL  REPLIC.                                                                              93   47  53  86  161  64  64   37  38                             MEAN WTS                                                                              PHBE2 101  100 105 99  99   101 102  100 94                             MEAN WTS                                                                              PHR03 101  104 95  101 101  102 100  100 104                                   DIFF  0    4   10  3   2    1   2    0   10                             __________________________________________________________________________

                                      TABLE 4B                                     __________________________________________________________________________     Average Inbred By Tester Performance Comparing PHBE2 to PHM46 Crossed To       The Same                                                                       Inbred Testers And Grown in The Same Experiments. All Values Are               Expressed As Percent Of                                                        The Experiment Mean Except Predicted RM, Selection Index, and Yield            (BU/ACR)                                                                                        SEL BU          GDU PRM STK RT  STA                                  INBRED                                                                               PRM IND ACR YLD MST SHD SHD LDG LDG GRN                           __________________________________________________________________________     TOTAL  REPLIC.                                                                              114 124 126 126 126 39  24  96  39  67                            MEAN WTS                                                                              PHBE2 111 105 151 103 95  101 113 97  102 121                           MEAN WTS                                                                              PHN46 111 104 149 102 94  100 112 101 103 106                                  DIFF      1   2   1   0   1   1   3   1   14                            __________________________________________________________________________                  TST  GRN SDG EST STK  PLT EAR  DRP BRT                                   INBRED                                                                               WTA  APP VGR CNT CNT  HT  HT   EAR STK                            __________________________________________________________________________     TOTAL  REPLIC.                                                                              89   50  53  74  147  58  58   43  27                             MEAN WTS                                                                              PHBE2 101  103 113 100 100  102 101  99  93                             MEAN WTS                                                                              PHN46 100  101 112 101 99   97  98   100 98                                    DIFF  2    2   1   1   1    5   3    1   5                              __________________________________________________________________________

DEPOSITS

Applicant has made a deposit of at least 2500 seeds of inbred Corn Line PHBE2 with the American Type Culture Collection (ATCC), Rockville. Md. 20852 USA, ATCC Deposit No. 97509. The seeds deposited with the ATCC on Apr. 11, 1996 were taken from the deposit maintained by Pioneer Hi-Bred International, Inc., 700 Capital Square, 400 Locust Street, Des Moines, Iowa 50309-2340 since prior to the filing date of this application. This deposit of the Inbred Corn Line PHBE2 will be maintained in the ATCC depository, which is a public depository, for a period of 30 years, or 5 years after the most recent request, or for the effective life of the patent, whichever is longer, and will be replaced if it becomes nonviable during that period. Additionally, Applicant has satisfied all the requirements of 37 C.F.R. §§1.801-1.809, including providing an indication of the viability of the sample. Applicant imposes no restrictions on the availability of the deposited material from the ATCC; however. Applicant has no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce. Applicant does not waive any infringement of its rights granted under this patent or under the Plant Variety Protection Act (7 USC 2321 et seq.). Applicant has applied for U.S. Plant Variety Protection of PHBE2 under Application No. 9500200.

The foregoing invention has been described in detail by way of illustration and example for purposes of clarity and understanding. However, it will be obvious that certain changes and modifications such as single gene modifications and mutations, somoclonal variants, variant individuals selected from large populations of the plants of the instant inbred and the like may be practiced within the scope of the invention, as limited only by the scope of the appended claims. 

What is claimed is:
 1. Seed of maize inbred line designation PHAP8 and having ATCC Accession No.
 97509. 2. A maize plant produced by the seed of claim 1 and its plant parts.
 3. Pollen of the plant of claim
 2. 4. An ovule of the plant of claim
 2. 5. A maize plant having all the physiological and morphological characteristics of the maize plant named PHBE2, the seed of which have been deposited under ATCC Accession No.
 97509. 6. A male sterile maize plant otherwise having all the physiological and morphological characteristics of the maize plant named PHBE2, the seed of which have been deposited under ATCC Accession No.
 97509. 7. A tissue culture of regenerable cells of an inbred maize plant PHBE2, wherein the tissue culture regenerates plant capable of expressing all the morphological and physiological characteristics of the maize plant named PHBE2, the seed of which have been deposited under ATCC Accession No.
 97509. 8. The tissue culture of claim 7, the tissue culture selected from the group consisting of leaves, pollen, embryos, roots, root tips, anthers, silk, flowers, kernels, ears, cobs, husks, stalks, cells and protoplasts thereof.
 9. A maize plant regenerated from the tissue culture of claim 7, capable of expressing all the morphological and physiological characteristics of inbred PHBE2 the seed of which have been deposited under ATCC Accession No.
 97509. 10. A method for producing first generation (F₁) hybrid maize seed comprising crossing a first inbred parent maize plant with a second inbred parent maize plant and harvesting the resultant first generation (F₁) hybrid maize seed, wherein said first or second parent maize plant is the maize plant of claim
 2. 11. The method of claim 10 wherein inbred maize plant PHBE2, the seed of which have been deposited under ATCC Accession No. 97509, is the female parent.
 12. The method of claim 10 wherein inbred maize plant PHBE2, the seed of which have been deposited under ATCC Accession No. 97509, is the male parent.
 13. An F₁ hybrid seed or plant, or parts thereof, produced by crossing an inbred maize plant according to claim 2 with another, different maize plant. 